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May 05, 2024
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2022-2023 Catalog [ARCHIVED CATALOG]
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BIOT 251 - Quantitative Real-Time Polymerase Chain Reaction and Related Applications
PREREQUISITES: BIOT 211 - Analytic Methods in Biotechnology I and CHEM 105 - General Chemistry I ; or Program Chair Approval.
PROGRAM: Biotechnology
CREDIT HOURS MIN: 5
LECTURE HOURS MIN: 2
LAB HOURS MIN: 6
DATE OF LAST REVISION: Fall, 2019
Quantitative real-time polymerase chain reaction (qPCR) is considered as the gold standard methods in molecular biotechnology and related applications. QPCR has a wide dynamic range with extreme sensitivity and specificity to detect and quantify few copies of target nucleic acid. This course will cover all pre and post PCR steps, from nucleic acid extraction to data analysis. This course is considered as a multidisciplinary course because it will cover different topics and students will acquire several skills. The course will cover the different chemistries of qPCR as well as the instrument hardware structures such as heating, cooling system and excitation detection system. The relationship between instrument hardware and reaction efficiency and accuracy will be discussed in detail. Furthermore, part of computational biology of bioinformatics, instrument and data analysis software will be covered. Moreover, special pipetting and samples handling skills will be acquired by students. At the end of the course, students will be professionally able to establish a qPCR lab and to decide what instrument and chemistry to be used, students will be able to read commercially available kits and apply it in the lab with high reproducibility and professionally analyze results.
MAJOR COURSE LEARNING OBJECTIVES: Upon successful completion of this course the student will be expected to:
- Gain knowledge related to the principles of conventional and quantitative real-time PCR.
- Distinguish the different types of quantitative real-time PCR chemistries.
- Determine the different types of quantitative real-time PCR platform.
- Establish and set up a quantitative real-time PCR lab and to determine the equipment needed.
- Utilize bioinformatics to identify the sequence of marker genes and the cDNA sequence.
- Design, check-up and manipulate different types of primers and probes.
- Review and develop Experimental design, and plate and protocol setup for DNA and gene expression quantification and standard curve generation.
- Design, setup, run and analyze results for marker gene quantification by nonspecific quantitative real-time PCR with melt curve (DNA template, primers, and master mix preparation).
- Design, setup, run and analyze results for marker gene quantification by specific quantitative real-time PCR (DNA template, primers and probe, and master mix preparation).
- Design, setup, run and analyze results for gene expression quantification and profiling by nonspecific quantitative real-time PCR with melt curve (RNA/cDNA templates, primers, and master mix preparation).
- Design, setup, run and analyze results for gene expression quantification by specific quantitative real-time PCR (RNA/cDNA templates, primers and probe, and master mix preparation).
- Detect (quantitatively) genetically modified organisms (GMO) by quantitative real-time PCR.
- Quantify microRNA by quantitative real-time PCR (specific microRNA and microRNA profiling).
- Identify genotype single-nucleotide polymorphisms (SNPs) by specific and nonspecific (melt curve) quantitative real-time PCR.
- Utilize commercially available kits.
- Develop a quantitative real-time PCR kit.
COURSE CONTENT: Topical areas of study include -
- Conventional vs quantitative real-time PCR
- Real-time PCR chemistries
- Real-time PCR platforms
- Bioinformatics, primers and probes design and analysis
- Quantitative real-time PCR and reverse transcription quantitative real-time PCR reaction setup
- Experiment design and data analysis
- Applications of PCR
GRADING POLICY
| A |
90-100 |
| B |
80-89 |
| C |
70-79 |
| D |
60-69 |
| F |
0-59 |
Course Addendum - Syllabus (Click to expand)
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